Heparanase and Syndecan-4 Are Involved in Low Molecular Weight Fucoidan-Induced Angiogenesis

Induction of angiogenesis is a potential treatment for chronic ischemia. Low molecular weight fucoidan (LMWF), the sulfated polysaccharide from brown seaweeds, has been shown to promote revascularization in a rat limb ischemia, increasing angiogenesis in vivo. We investigated the potential role of two heparan sulfate (HS) metabolism enzymes, exostosin-2 (EXT2) and heparanase (HPSE), and of two HS-membrane proteoglycans, syndecan-1 and -4 (SDC-1 and SDC-4), in LMWF induced angiogenesis. Our results showed that LMWF increases human vascular endothelial cell (HUVEC) migration and angiogenesis in vitro. We report that the expression and activity of the HS-degrading HPSE was increased after LMWF treatment. The phenotypic tests of LMWF-treated and EXT2- or HPSE-siRNA-transfected cells indicated that EXT2 or HPSE expression significantly affect the proangiogenic potential of LMWF. In addition, LMWF increased SDC-1, but decreased SDC-4 expressions. The effect of LMWF depends on SDC-4 expression. Silencing EXT2 or HPSE leads to an increased expression of SDC-4, providing the evidence that EXT2 and HPSE regulate the SDC-4 expression. Altogether, these data indicate that EXT2, HPSE, and SDC-4 are involved in the proangiogenic effects of LMWF, suggesting that the HS metabolism changes linked to LMWF-induced angiogenesis offer the opportunity for new therapeutic strategies of ischemic diseases.     

Continuous positive airway pressure administered via face mask in tranquilized dogs

Objective – To evaluate the tolerance of a continuous positive airway pressure (CPAP) mask in tranquilized dogs and compare PaO₂ in arterial blood in dogs receiving oxygen with a regular face mask or CPAP mask set to maintain a pressure of 2.5 or 5 cm H₂O. Design – Prospective, randomized clinical study. Setting – University teaching hospital. Animals – Sixteen client‐owned dogs without evidence of cardiopulmonary disease were studied. Interventions – Eight animals were randomly assigned to each of 2 treatment groups: group A received 2.5 cm H₂O CPAP and group B received 5 cm H₂O CPAP after first receiving oxygen (5 L/min) by a regular face mask. Animals were tranquilized with acepromazine 0.05 mg/kg, IV and morphine 0.2 mg/kg, IM. An arterial catheter was then placed to facilitate blood sampling for pHa, PaO₂, and PaCO₂ determinations before and after treatments. Direct mean arterial pressure, heart rate, respiratory rate, and temperature were also recorded after each treatment. Measurements and Main Results – CPAP administration was well tolerated by all animals. The mean arterial pressure, heart rate, respiratory rate, temperature, PaCO₂, and pHa, did not differ at any time point between groups. Differences were seen in oxygenation; in group A, PaO₂ significantly increased from a mean of 288.3±47.5 mm Hg with a standard mask to a mean of 390.3±65.5 mm Hg with the CPAP mask and in group B, PaO₂ increased similarly from 325.0±70.5 to 425.2±63.4 mm Hg (P<0.05); no differences were detected between the 2 CPAP treatments. Conclusions – In healthy tranquilized dogs noninvasive CPAP is well tolerated and increases PaO₂ above values obtained when using a regular face mask.     

Implications of the number of years assessment on recommendation of common bean cultivars

In the common bean crop in Brazil, the requirement of the value for cultivation and use trials is that these experiments must be conducted over two years in three locations per region. Information in regard to the necessary number of years to ensure precision in cultivar recommendation and the influence of evaluated years in the estimation of the GE interaction are still scarce. Using grain yield of five check varieties assessed over 11 years in three sowing seasons per year, the aims of this study are as follows: to measure the relative contribution of the GE interaction, evaluating the implication of the number of years in the estimates of the GE interaction, and infer how many years are needed to ensure precision in cultivar recommendation. For instances, analysis of variance was carried out involving all environments and also combinations of years. The results showed that the GE interaction was greater than all other cross‐effects involving lines. The use of at least two years allows good coincidence in cultivar recommendation compared to the whole period. Increasing the evaluation time is a good strategy, especially when it is difficult to grow three different sowing seasons.     

Suitability of soil microbial parameters as indicators of heavy metal pollution

Several microbial parameters (microbial biomass, respiration, dehydrogenase, phosphatase, sulphatase, glucosidase, protease and urease activities) were measured in soils from five sites located in urban green areas close to roads differing in traffic density. Our aims were to evaluate the suitability of such parameters as field biomarkers of stress induced by heavy metal pollution, and to compare results obtained by single microbial parameters with results given by an index expressing the average microbial (AME) response of the microbial community. Data showed that all parameters were significantly reduced in the sites characterized by the highest load of metals in soil. Dehydrogenase, sulphatase, glucosidase activities and respiration, declined exponentially with increasing metal concentration, whereas phosphatase activity and AME decreased following a sigmoidal type relationship. In contrast, protease, urease and microbial biomass were not significantly correlated with soil metal concentration. Microbial parameters differed both in sensitivity to critical metal concentrations and in the rate of decline at increasing metal loads in soil. Due to the complex interplay of chemical, physical and biological factors which influence microbial activities and biomass, the proposed index (AME) appeared more suitable than single microbial parameters for a biomonitoring study of this type.     

Supercritical extraction of essential oil from aniseed (Pimpinella anisum L) using CO2: solubility,kinetics, and composition data

Supercritical fluid extraction (SFE) from aniseed using carbon dioxide was performed at 30 degrees C and pressures of 80-180 bar. The chemical composition of the SFE extract was determined by GC-MS; the quantitative analysis was done by GC-FID and TLC. The total amount of extractable substances or global yield (mass of extract/mass of feed) for the SFE process varied from 3.13 to 10.67% (mass). The solubilities of the anise essential oil in CO(2) were 0.0110, 0.0277, 0.0143, and 0.0182 kg of solute/kg of CO(2) at 80, 100, 140, and 180 bar, respectively. The major compounds identified and quantified in the extracts were anethole ( approximately 90%), gamma-himachalene (2-4%), p-anisaldehyde (<1%), methylchavicol (0.9-1.5%), cis-pseudoisoeugenyl 2-methylbutyrate ( approximately 3%), and trans-pseudoisoeugenyl 2-methylbutyrate ( approximately 1.3%). The Sovová model described quite well the experimental overall extraction curves.     

Reaction of <Emphasis Type="Italic">Phaseolus</Emphasis> spp. genotypes to ashy stem blight caused by <Emphasis Type="Italic">Macrophomina phaseolina</Emphasis>

Ashy stem blight (ASB) caused by Macrophomina phaseolina (Tassi) Goidanich (Mp) is a devastating seed-transmitted disease in common bean in the tropics. The identification of resistant cultivars throughout the cropping season contributes to disease management. Resistance is found in the primary and tertiary gene pools. Our objectives were to determine (1) the reaction of Phaseolus spp. genotypes to two Mp isolates at vegetative and reproductive stages, (2) the area under disease progress curve (AUDPC), and (3) resistant plants per genotype at harvest. Twenty-three genotypes from different origins were planted in the greenhouse in 2016 and 2017. One less-aggressive Mp (PRJD16) and one more-aggressive (PRI16) isolate were inoculated one and three times, respectively, by the cut-stem method. ‘Beníquez’, ‘Othello’, and ‘Verano’ were highly susceptible (mean scores >8.0, and AUDPC values from 264.6 to 300.8) to both isolates. BAT 477 and NY6020-4 were intermediate (5.6 and 6.2; AUDPC: 161.6 and 187.1) to PRJD16 and susceptible (7.4 and 8.2; AUDPC: 209.4 and 235.1) to PRI16. Resistant genotypes (mean scores ≤3) were not identified in this study. However, A 195, ‘Badillo’, and ‘PC 50’ possessed lower mean scores (4.3–5.4) and AUDPC values (126.4–150.9) to both isolates. Furthermore, A 195 had the highest percentage of resistant plants (55.6%) followed by PC 50, I9365-31, and PI 321637 (27.8%) to PRJD16 at harvest. Thus, the identification of resistant parents across Phaseolus species is necessary to increase the levels of ASB resistance in common bean cultivars throughout the entire cropping season.     

Extraction of bacteria from soil: Efficiency of shaking or ultrasonication as indicated by direct counts and autoradiography

Blending, shaking with a mechanical wrist-action shaker or ultrasonication were compared for removing bacteria from a sandy soil and a silt loam soil. Bacteria were counted microscopically after staining with acridine orange. There was no significant difference in the numbers of bacteria released from the sandy soil by the different treatments but ultrasonication extracted more bacteria from the silt loam. Highest recoveries of bacteria were obtained with a combination of Tris buffer as extractant, and 25% Ringer's solution as diluent. Ultrasonication of an irradiated sandy soil containing added Bacillus cereus removed 97% of the bacteria from the sand grains, but lysed some cells. Members of the indigenous soil population were more firmly held by the soil than added bacteria. Optimum conditions for recovering soil bacteria by ultrasonication were either 10.5 μm for 30s or 5 μm for 2 min. With the natural population, the numbers of bacteria and the proportion of metabolizing bacteria increased a little after ultrasonication as shown by labelling cells with ³H-glucose and detecting uptake by autoradiography. However, three isolates of bacteria tested showed varying amounts of injury after ultrasonication, with 5 μm for 2 min causing more damage than 10.5 μm for 30s. Ultrasonication was a more efficient means of extracting the natural bacterial population of soil than shaking but the time and amplitude must be adjusted to avoid injury to cultured bacterial cells.     

Echocardiography in the chinchilla

Heart disease has been described in the chinchilla and, with increasing popularity as a pet, the demand for diagnostic evaluation and treatment has increased. The goal of this study was to determine reference values for echocardiographic measurements in chinchillas and the effect of anesthesia on these measurements. Seventeen clinically healthy adult chinchillas were studied. All animals were anesthetized with isoflurane by mask. Standard echocardiographic views were used. A difference was seen in the echocardiographic measurements for left ventricular systolic dimension, fractional shortening, aortic (Ao) diameter, left atrial (LA) diameter, ratio of LA diameter to Ao diameter, and peak flow velocities and ejection times for Ao and pulmonary artery flows between awake and anesthetized chinchillas.     

Competition Effects Among Isolates of Fusarium culmorum Differing in Aggressiveness and Mycotoxin Production on Heads of Winter Rye

Fusarium culmorum is a phytopathogenic, toxigenic fungus causing seedling diseases, foot rot and head blight of cereals. For estimating competition effects in mixtures of two single-spore isolates, two winter rye single crosses were tested with either four isolates individually or four 1 : 1 mixtures of the same isolates in six field environments. Two isolates (FC46, FC64) were highly aggressive deoxynivalenol (DON) and 3-acetyl DON-producers, the other two (FC30, FC71) were medium aggressive nivalenol-producers. Rye heads were inoculated during flowering with conidia of pairs of isolates expressing similar (FC46 + FC64, FC30 + FC71) or contrary (FC46 + FC71, FC30 + FC64) levels of aggressiveness and similar or different concentrations and chemotypes of mycotoxins, respectively. Head blight rating and yield components relative to the non-inoculated plots were recorded as aggressiveness traits. Additionally, mycotoxin concentrations were measured in the rye grain. Random pathogen samples were re-isolated from heads at the onset of symptom development and analysed by molecular markers (RAPD–PCR) in one environment. Aggressiveness of the isolate mixtures was significantly lower than that of the isolates applied individually on both rye genotypes. Similarly, mycotoxin concentrations were significantly lower in the mixtures in seven out of eleven comparisons. Among the re-isolates, the component genotypes of a mixture significantly deviated from the inoculated 1 : 1 ratio when a particular isolate (FC46) was present in the mixture. This isolate displayed a superior competitive ability irrespective of the aggressiveness or mycotoxin profile of the mixing partner illustrating that pathogenic fitness is caused by additional factors that have not, as yet, been identified.     

Content of Nutrient and Antinutrient in Edible Flowers of Wild Plants in Mexico

Nutrient and antinutritional/toxic factors present in some edible flowers consumed in Mexico were determined. The edible flowers were: Agave salmiana, Aloe vera, Arbutus xalapensis, Cucurbita pepo (cultivated), Erythrina americana, Erythrina caribaea, Euphorbia radians benth and Yucca filifera. The nutrient content in the flowers studied is similar to that of the edible leaves and flowers studied mainly in Africa. The moisture content of the flowers varied from 860 to 932 g kg⁻¹. Crude protein (CP) was between 113 to 275 g kg⁻¹ DM, crude fiber, 104 to 177 g kg⁻¹ DM and the nitrogen free extract, between 425 to 667 g kg⁻¹ DM. The highest chemical score (CS) was found in E. americana and A. salmiana; in five samples the limiting amino acid was lysine, and in three of them it was tryptophan. Trypsin inhibitors and hemaglutinnins had a very low concentration. Alkaloids were present in both the Erythrina species and the saponins in A. salmiana and Y. filifera. Cyanogenic glucosides were not found in the studied flowers. The traditional process of preparing these specific flowers before consumption is by cooking them and discarding the broth; in this way the toxic substances are diminished or eliminated. These edible flowers from wild plants consumed in local areas of the country play an important role in the diet of the people at least during the short time of the season where they are blooming.